Green tea flower formulation for weight management

ABSTRACT

The invention is directed to a composition formulated from green tea flower and methods for making and using the composition in the management of body weight.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of application Ser. No. 16/580,982,filed Sep. 24, 2019, which claims the benefit of provisional applicationNo. 62/736,399, filed Sep. 25, 2018. The entire contents of theseapplications are incorporated herein by reference for all purposes.

FIELD OF THE INVENTION

The invention is in the field of preparations for weight management.

BACKGROUND

Green tea (Camelia sinensis L) prepared from tea leaves is one of themost widely consumed non-alcoholic beverages in the world. It hasattained significant attention owing to its benefits against array ofhealth problems such as obesity, diabetes mellitus, cardiovasculardisorders, and cancer. These beneficial effects have been partlyattributed to its chemical ingredients, includingepigallocatechinin-3-gallate, epicatechinin-3-gallate,epigallocatechinin, and epicatechinin to name a few. The presentdisclosure and accompanying data enable a novel application of greentea, including the use of green tea flowers in the formulation of acomposition for weight management.

SUMMARY OF THE INVENTION

It is an object of the invention to provide a composition formulatedfrom green tea flower for the management of body weight.

In some aspects, the composition comprises green tea flower extract.

It is a further object of the invention to provide a method ofmodulating body weight comprising administering to a patient in needthereof a composition comprising green tea flower extract.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts graphs showing the free radical scavenging activity of anembodiment the inventive composition.

FIG. 2 depicts graphs showing the inhibitory effect of an embodiment ofthe inventive composition on pancreatic lipase and alpha amylase.

FIG. 3 is a graph showing the effect of an embodiment of the inventivecomposition on the viability of 3T3-L1 preadipocytes.

FIG. 4A depicts micrographs showing the anti-adipogenic effect of anembodiment of the inventive composition as determined by Oil Red Ostaining.

FIG. 4B is a graph showing the anti-adipogenic effect of an embodimentof the inventive composition as determined by lipid content.

FIG. 5 is a Western blot showing the effect of an embodiment of theinventive composition on the expression of PPARγ and C/EBPα on day 8.

DEFINITIONS

As used herein, the term “about” means the quantity, level, value,number, frequency, percentage, dimension, size, amount, weight or lengththat is referenced, or that varies (plus or minus) by as much as 30%,25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of thereferenced quantity, level, value, number, frequency, percentage,dimension, size, amount, weight or length.

Throughout this specification, unless the context requires otherwise,the words “comprise,” “comprises,” and “comprising” will be understoodto refer to the inclusion of a stated step or element, or group of stepsor elements, but not the exclusion of any other step or element or groupof steps or elements.

As used herein, the term “increase” refers to any measurable increase ina parameter relative to control conditions.

As used herein, the term “modulate” refers to the act of effecting analteration or change in the natural state of a parameter or condition.For example, “modulate” can refer to an increase, decrease, theinhibition of, or preventing a change in, the body weight or body fat ofa patient.

As used herein, the term “reduce” refers to any measurable decrease in aparameter relative to control conditions.

As used herein, the terms “subject,” “patient,” “individual” and liketerms are used interchangeably and refer to, except where indicated,mammals such as humans and non-human primates, as well as livestock andcompanion and laboratory research animals.

DETAILED DESCRIPTION

The invention generally relates to the management of body weight. Moreparticularly, the invention relates to a composition formulated fromgreen tea flower extract and a method of its use and manufacture in themanagement of body weight.

The inventor surprisingly discovered a composition of green flower teaextract can be highly effective in managing a patient's body weight. Asexemplified by the present disclosure, the composition has demonstratedeffects on many aspects of the physiological regulation of body weight.In particular, the inventor has shown that the composition can inhibitpancreatic lipase and alpha amylase and impact adipocyte development.The inventor further demonstrated that the composition can impact theexpression and activity of proteins involved in weight metabolism,including the transcription factors PPARγ and C/EBPα which are knownadipogenesis regulators.

In some aspects, the invention provides a composition. The compositioncan comprise green tea flower extract. The composition can comprisegreen tea extract and one or more excipient or one or more carriers. Insome aspects, the composition is free of an extract of green tea leaves.The composition can comprise green tea extract and one or more excipientand one or more carriers. The excipient and/or carrier can be selectedon the basis of compatibility with green tea flower extract and theproperties of the desired dosage form. Suitable excipients include, butare not limited to, binders, fillers, bulking agents, flowaids/glidents, disintegrants, lubricants, stabilizers, surfactants, andthe like. The excipient can be selected from the group consisting ofbinders, fillers, bulking agents, flow aids/glidents, disintegrants,lubricants, stabilizers, surfactants, and combinations thereof. Suitableexcipients and carriers for use with the composition include, but arenot limited to, those disclosed in: Remington: The Science and Practiceof Pharmacy, 19^(th) Ed (Easton, Pa.: Mack Publishing Company, 1995);Hoover, John E., Remington's Pharmaceutical Sciences, (Easton, Pa.: MackPublishing Co 1975); Liberman, H. A. and Lachman, L., Eds.,Pharmaceutical Dosage Forms (New York, N.Y.: Marcel Decker 1980); andPharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed(Lippincott Williams & Wilkins 1999). The entire contents of thesepublications are incorporated herein by reference for all purposes.

In some aspects of the invention, the composition employs controlled,sustained, or extended release formulations known collectively as“modified release” formulations. The composition can be administered bymodified release systems or by delivery devices known to those skilledin the art. Suitable examples of such systems and delivery devicesinclude, but are not limited to, those described in the followingpatents, the entire disclosures of which are incorporated herein byreference for all purposes: U.S. Pat. Nos. 3,845,770; 3,916,899;3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767;5,120,548; 5,073,543; 5,639,476; 5,354,556; and 5,733,566. Dosage formsfor the composition can be used to provide modified release of one ormore active ingredients using, for example, hydropropylmethyl cellulose,other polymer matrices, gels, permeable membranes, osmotic systems,multilayer coatings, microparticles, liposomes, microspheres, orcombinations thereof

At least one aspect of the invention concerns the dosage form of thecomposition. The composition can be in the form of a powder, liquid,pill, tablet, pellet, capsule, thin film, solution, spray, syrup,linctus, lozenge, pastille, chewing gum, paste, vapor, suspension,emulsion, ointment, cream, lotion, liniment, gel, drop, topical patch,buccal patch, bead, gummy, gel, sol, injection, or combinations thereof.The composition can be formulated for oral administration. Thecomposition can be combined with vitamins, minerals, amino acids,proteins, extracts, carbohydrates, lipids, fatty acids, caffeine,flavorings, sweeteners, preservatives, or combinations thereof.

In some aspects of the invention, the composition is combined with afood, snack, nutritional supplement, dietary supplement, foodsupplement, or beverage. The food, snack, nutritional supplement,dietary supplement, food supplement, or beverage can have reducedcalorie content. For example, the composition can be provided as a meansfor managing body weight wherein the composition is formulated toprovide a dietetic food, a dietetic snack, a dietetic nutritionalsupplement, a dietetic dietary supplement, a dietetic food supplement,or a dietetic beverage. The composition can be combined with dieteticsnacks such as bars, chips, chews, gels, candies, chocolates, cakes,cookies and other pastries, wafers, crackers, ice cream, and the like.

The composition can be provided in bulk quantities for the industrialmanufacture of the products and dosage forms described herein. Forexample, the composition can be provided in bulk quantities as a powderor liquid. Bulk quantities of the composition can be packaged, storedand/or distributed in drums, bags, boxes, and other containers which areconfigured to prevent or inhibit the oxidation of one or more activecomponents of the composition.

In some aspects, the invention provides a method of modulating bodyweight in a patient. The method can be practiced by administering to thepatient the composition of the present invention. In some preferredembodiments, the composition is administered in an effective amount. Thecomposition can be administered to the patient to reduce body weight, orto inhibit or prevent weight gain in the patient. For example, thecomposition can be administered to maintain a desired body weight in thepatient. Administering the composition can reduce body fat in thepatient, or inhibit or prevent an increase in body fat in the patient.The patient can be obese, overweight or have a normal body weight. Theterms “obese” and “obesity” as used herein refer to a patient having abody mass index of 30 or higher. The term “overweight” as used hereinrefers to a patient having a body mass index of 25 to 29.9. The term“healthy weight” as used herein refers to a patient having a body massindex of 18.5 to 24.9. In at least one aspect of the invention, thepatient is human.

Without being limited to any particular theory or mechanism,administering the composition can impart its effect by, among othermeans, inhibiting one or more forms of lipase in the patient. Lipases(e.g. triacylglycerol hydrolase E.C. 3.1.1.3) are enzymes that catalyzethe hydrolysis of ester bonds of triacylglycerols (fats and oils) toproduce free fatty acids such as diacylglycerols, monoglycerols andglycerol. In the small intestine of mammals, the digestion of dietarytriacylglycerols (TAG) is essentially due to the action of pancreaticlipase. The end products of TAG digestion are absorbed by the body andare responsible for the development of weight gain and obesity.Therefore, if the hydrolysis of TAG and its movement from the intestinallumen into the body is stopped or minimized, the prevalence of obesitycan be reduced. The composition can inhibit pancreatic lipase,pancreatic lipase related protein 2, hepatic lipase, endothelial lipase,lipoprotein lipase, or combinations thereof. In one non-limitingembodiment, administering the composition inhibits pancreatic lipase inthe patient.

At least one aspect of the invention concerns the dosage at which thecomposition is administered to the patient. The composition can beadministered at a dose of between about 5 mg/day and about 500 mg/day.The composition can be administered at a dose between about 20 mg/dayand about 1 mg/day. The composition can be administered at a dose ofabout 20 mg/day, about 21 mg/day, about 22 mg/day, about 23 mg/day,about 24 mg/day, about 25 mg/day, about 26 mg/day, about 27 mg/day,about 27 mg/day, about 28 mg/day, about 29 mg/day, about 30 mg/day,about 31 mg/day, about 32 mg/day, about 33 mg/day, about 34 mg/day,about 35 mg/day, about 40 mg/day, about 45 mg/day, about 50 mg/day,about 100mg/day, about 150 mg/day, about 200 mg/day, about 250 mg/day,about 300 mg/day, about 350 mg/day, about 400 mg/day, about 450 mg/day,or about 500 mg/day, as well as any dosage intervening thesespecifically disclosed amounts. The composition can be administered at adosage of between about 400 mg/day and about 500 mg/day, between about300 mg/day and about 400 mg/day, between about 200 mg/day and about 300mg/day, between about 100 mg/day and about 200 mg/day, or about 20mg/day and about 100 mg/day. It is contemplated that the composition canbe administered at any dosage that intervenes the dosages called out inthis specification.

In some aspects, the dosage of the composition is determined by the bodyweight of the patient. The composition can be administered between about5 mg/kg b.w. and about 500 mg/kg b.w. The composition can beadministered at about 5 mg/kg b.w., about 10 mg/kg b.w., about 20 mg/kgb.w., about 30 mg/kg b.w., about 40 mg/kg b.w., about 50 mg/kg b.w.,about 60 mg/kg b.w., about 70 mg/kg b.w., about 80 mg/kg b.w., about 100mg/kg b.w., about 120 mg/kg b.w., about 140 mg/kg b.w., about 160 mg/kgb.w., about 180 mg/kg b.w., about 200 mg/kg b.w., about 220 mg/kg b.w.,about 240 mg/kg b.w., about 260 mg/kg b.w., about 280 mg/kg b.w., about300 mg/kg b.w., about 320 mg/kg b.w., about 340 mg/kg b.w., about 360mg/kg b.w., about 380 mg/kg b.w., about 400 mg/kg b.w., about 420 mg/kgb.w., about 440 mg/kg b.w., about 460 mg/kg b.w., about 480 mg/kg b.w.,or about 500 mg/kg b.w. In one non-limiting embodiment of the invention,the composition is administered at about 150 mg/kg b.w. The compositioncan be administered, one, two, three, four, five or more times. Thecomposition can be administered daily, weekly, monthly, or combinationsthereof. The composition can be administered one, two, three, four,five, six or more times per day, per week, or per month. One skilled inthe art will appreciate that the administration of the composition canbe adjusted according to the patient's response to the treatment and thebody weight desired by the patient or attending physician.

At least one aspect of the invention concerns the administration routeof the composition. The composition can be administered systemically.Suitable administration routes for the composition include, but are notlimited to, auricular, buccal, conjunctival, cutaneous, dental,endocervical, endosinusal, endotracheal, enteral, epidural,extra-amniotic, interstitial, intra-abdominal, intra-amniotic,intra-arterial, intra-articular, intrabiliary, intrabronchial,intrabursal, intracardiac, intracartilaginous, intracaudal,intracavernous, intracavitary, intracerebral, intraci sternal,intracorneal, intracoronal dental, intracoronary, intracorporuscavernosum, intradermal, intradiscal, intraductal, intraduodenal,intradural, intraepidermal, intraesophageal, intragastric, intravaginal,intraileal, intralesional, intraluminal, intralymphatic, intramedullary,intrameningeal, intramuscular, intraocular, intraovarian,intrapericardial, intraperitoneal, intrapleural, intraprostatic,intrapulmonary, intrasinal, intraspinal, intrasynovial, intratendinous,intratesticular, intrathecal, intrathoracic, intratubular, intratumor,intratympanic, intrauterine, intravascular, intravenous, intravenousbolus, intravenous drip, intraventricular, intravitreal, laryngeal,nasal, nasogastric, ophthalmic, oral, oropharyngeal, parentera,percutaneous, periarticular, peridural, perineural, periodontal, rectal,inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival,subcutaneous, sublingual, submucosal, topical, transdermal,transmucosal, transplacental, transtracheal, transtympanic, ureteral,urethral, vaginal, or combinations thereof In a preferred embodiment,the composition is administered orally.

In at least one aspect, the invention provides a method for making thecomposition. The composition can be made by providing green tea flowers,and extracting one or more constituents effective in modulating bodyweight in the subject. The green tea flowers for making the compositioncan be flowers from Camelia sinensis. The green tea flowers can befresh, dried, or a combination thereof. The extract can be made usingany suitable process, including, without limitation, solvent extraction,extrusion, or a combination thereof. Suitable solvents for obtainingextracts for formulating the composition include, but are not limitedto, aqueous solvents, alcohol-based solvents, supercritical fluids,polar organic solvents (such as acetone and methylethyl ketone), orcombinations thereof. Non-limiting examples of alcohol-based solventsinclude, but are not limited to, ethanol, isopropyl alcohol, methanol,and combinations thereof. The supercritical fluid can be, but is notnecessarily limited to, carbon dioxide.

The present disclosure is further described in the light of thefollowing example which is set forth for illustration purposes only andis not to be construed as limiting the scope of the present invention.

EXAMPLE Sample Preparation

A hydro alcoholic extract of green tea flower was prepared and dissolvedin water at the appropriate concentrations.

DPPH Free Radical Scavenging Activity

DPPH Free radical scavenging activity of green tea flower extract (GTFE)was carried out by adopting the protocol described by Braca et al.,2001.

Superoxide Anion Scavenging Activity

Superoxide anion scavenging activity of GTFE was carried out by theprocedure described by McCord and Fridovich, 1969.

Nitric Oxide Scavenging Activity

Nitric oxide generated from sodium nitroprusside was measured by theGriess reagent by adopting the method described by Marcocci et al. 1994.

Anti-Lipid Peroxidation Assay (TBARS)

A modified thiobarbituric acid reactive species (TBARS) assay was usedto measure the lipid peroxide formed using egg yolk homogenates aslipid-rich media, as described by Ruberto et al., 2000.

Pancreatic Lipase Activity

Pancreatic lipase activity of GTFE was measured by adopting the methoddescribed by Kim Y S et al., 2010 with minor modifications. Briefly anenzyme buffer was prepared by the addition of a solution of porcinepancreatic lipase (2.5 mg/ml in 10 mM MOPS (morpholinepropanesulphonicacid) and 1 mM EDTA, pH 6.8) to 169 μl of Tris buffer (100 mM Tris-HCland 5 mM CaCl2, pH 7.0). Subsequently, 20 μl of different concentrationsof GTFE was mixed with 20 μl of the enzyme buffer and incubated for 15min at 37° C. with 5 μl of the substrate solution (10 mM p-NPB(p-nitrophenyl butyrate) in dimethyl formamide). The enzymatic reactionswere allowed to stand for 30 min at 37° C. Lipase activity wasdetermined by measuring the hydrolysis of p-NPB to p-nitrophenol at 405nm using microplate reader.

α-Amylase Inhibition Assay

This assay was carried out using a modified procedure (McCue and Shetty,2004). A total of 250 μL of extract (μg/mL) was placed in a tube and 250μL of 0.02 M sodium phosphate buffer (pH 6.9) containing a-amylasesolution (0.5 mg/mL) was added. This solution was pre-incubated at 25°C. for 10 min, after which 250 μL of 1% starch solution in 0.02 M sodiumphosphate buffer (pH 6.9) was added at timed intervals and then furtherincubated at 25° C. for 10 min. The reaction was terminated by adding500 μL of dinitrosalicylic acid (DNS) reagent. The tubes were thenincubated in boiling water for 3 min and cooled to room temperature. Thereaction mixture was diluted with 5 mL distilled water and theabsorbance was measured at 540 nm using spectrophotometer. A control wasprepared using the same procedure replacing the GTFE with distilledwater. The α-amylase inhibitory activity was calculated as a percentageinhibition.

Cell Culture and Differentiation

3T3-L1 fibroblasts were cultured to confluence in Dulbecco's modifiedEagle medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS:BCO) and 1% penicillin-streptomycin (GIBCO) in a CO₂ incubator at 37° C.On day 2 post-confluence (designated as day 0), cells were induced todifferentiate with DMEM containing 10% fetal bovine serum (FBS; GIBCO),5μg/ml insulin (Sigma, St. Louis, MO, USA), 1 μmol/L dexamethasone(Sigma), and 0.05 mmol/L 3-isobutyl-1-methylxanthine (IBMX; Sigma) (MDIcocktail). After 2 days, the medium was replaced with DMEM supplementedwith 10% FBS and 5μg/ml insulin. The cells were subsequently re-fedevery 48 h with DMEM containing 10% FBS. To examine the anti-adipogeniceffect of GTFE, GTFE was added to the medium at different concentrations(10, 25, 50, and 100 μg/ml) during medium changes.

MTT Assay

3T3-L1 fibroblasts were seeded at a density of 1×10⁴ cells/well in96-well plates. The cells were treated with different concentrations ofGTFE for 48 h. After completion of the treatment, the cells wereincubated with 0.5 mg/ml MTT (3-4,5- dimethylthiazol-2-yl-2, 3-diphenyltetrazolium bromide) for 4 h at 37° C. The supernatants were carefullyaspirated and 100 μL of DMSO was added to dissolve the formazan crystalproduct. Absorbance was measured at 560 nm using a microplate reader.

Oil Red O Staining

Adipocyte cell monolayers were gently rinsed twice withphosphate-buffered saline (PBS), fixed in a with 4% paraformaldehyde-PBS solution for 1 h at room temperature, stained with 0.5% Oil Red Oisopropyl alcohol for 1 h, and then washed with distilled water. Thecells were checked by a bright-field optical microscope (HS-100,OPTICAL, China). The cells were eluted with isopropyl alcohol andabsorbance was measured at 520 nm with a microplate reader.

Western Blot Analysis

Cells were harvested by scraping in 120 μL lysis buffer (RIPA). Theywere then incubated on ice for 20 min and centrifuged at 13,000 rpm for15 min. The supernatant was then transferred to a fresh tube. Proteinconcentrations were determined. Equal concentrations of protein (100 μgper lane) were loaded in the wells of 6-12% polyacrylamide gels. Afterthe electrophoretic run, proteins on gels were transferred to apolyvinylidene difluoride membrane (Millipore, Marlborough, MA, USA) andincubated in 5% non-fat milk at room temperature. The membrane wasincubated with PPARγ (Abcam, 1:500), C/EBPα (Cell Signaling Technology,1:1000), and β-actin (Santa Cruz Biotechnology, 1:200) overnight at 4°C. The blots were incubated with horseradish peroxidase-conjugatedsecondary antibody (Santa Cruz Biotechnology) for 1 h. The blots weredeveloped by enhanced chemiluminescence (Santa Cruz Biotechnology).

Statistical Analysis

Each experiment was repeated three times and the data averaged forreporting. Data is given as the mean±standard error. Statisticalanalysis of data was determined by analysis of variance (ANOVA). Allstatistical tests were performed at the P<0.05 of significance.

Results

Reactive oxygen species (ROS), such as hydroxyl radicals, superoxideanions, and hydrogen peroxide can cause oxidative damage to cellularbiomolecules. High levels of reactive oxygen species are intricatelylinked to obesity and associated pathologies, notably insulin resistanceand type 2 diabetes. In this study, free radical scavenging activity ofthe composition of the invention was carried out using different invitro models. The results of DPPH, superoxide anion, nitric oxidescavenging properties of GTFE are shown in FIG. 1 .

DPPH Radical Scavenging Activity

GTFE showed concentration dependent DPPH radical scavenging activity(FIG. 1 ) with an IC50 value 40.9 μg/mL. The effect of antioxidants onDPPH radical scavenging was thought to be due to their hydrogen donatingability.

Superoxide Anion Scavenging Activity

The results of this study showed a concentration-dependent pattern forthe scavenging of superoxide anion radicals. GTFE distinctly showedscavenging ability and an IC 50 value found to be 50.7 μg/mL (FIG. 1 )

Nitric Oxide Scavenging Activity

GTFE inhibited the formation of nitric oxide from sodium nitroprside. Adose dependent pattern for nitric oxide scavenging potential wasexhibited by GTFE (FIG. 1 ). For scavenging nitric oxide radicals, GTFEshowed an IC50 of 43.7 μg/mL

Inhibition of Lipid Peroxidation

GTFE was analyzed for its ability to inhibit lipid peroxidation. Thegeneration of malondialdehyde (MDA) and related substances that reactwith thiobarbituric acid was found to be inhibited by GTFE (FIG. 1 ).The IC50 value of GTFE for lipid peroxidation was found to be 42.7μg/mL.

Effect on Pancreatic Lipase and Amylase Activity

FIG. 2 presents GTFE's effect on pancreatic lipase and a-amylaseactivity. GTFE showed a concentration dependent inhibitory activitytowards pancreatic lipase and α-amylase, the calculated IC50 value forpancreatic lipase and α-amylase activity were 120.42 and 198.7 μg/mLrespectively. The lipase activity in digestive system breaks downdietary fats and oils into free fatty acids. These free fatty acids areabsorbed into the blood stream and occur in circulation astriglycerides. The increased level of serum triglycerides provides amarker for altered lipid metabolism in the body. In the present study,GTFE showed inhibitory activity. Inhibition of the enzyme alpha-amylasedecreases the hydrolysis of starch into simple sugars. Sucrase cleavessucrose into glucose and fructose, the inhibition of which control theconversion of disaccharides into simple sugars. In the present studyGTFE inhibited α-amylase activity.

Effect on 3T3-L1 Cell Viability

MTT assay results showed reduction in viability of 3T-3L1 cells afterGTFE exposure in a dose dependent manner, showing less viability athigher extract concentrations. The IC50 value was determined to be ˜292μg/ml after 48 h of exposure (FIG. 3 ). For further study ondose-dependence, 150 μg/ml and 250 μg/ml doses of GTFE were taken.

Effects on Adipogenic Differentiation

In order to investigate the effects of GTFE on preadipocytedifferentiation, lipid accumulation was measured by Oil Red O stainingassay. As shown in FIGS. 4A and 4B, 250 μg/mL of GTFE suppressed lipidaccumulation in 3T3-L1 adipocytes at levels that were statisticallysignificant (p<0.05), showing that GTFE can inhibit adipogenesis in3T3-L1 cells. The use of differentiation of undifferentiated fibroblasts(pre-adipocytes) to mature adipocytes, which is termed adipogenesis, isa central area of obesity research. The main characteristics of cellularadipogenesis are continuous fat mobilization and subsequent cellmorphological changes in size and shape. During adipogenesis, fatdroplets accumulate in adipocytes and cells become moreinsulin-responsive. Also, gene expression of fat-related factors arechanged. In this study, the ability of the composition of the inventionto inhibit the mechanism of adipogenesis was observed.

Effects on PPARγ and C/EBPα

In order to investigate anti-adipogenesis, it was demonstrated that GTFEtreatment resulted in a dose-dependent suppression of the protein levelsof PPARγ and C/EBPα (FIG. 5 ). These results indicated that GTFEtreatment inhibited the expression of PPARγ and C/EBPα at the proteinlevel demonstrating that GTFE inhibited adipogenesis by suppressing thetranscriptional factor cascade. It has been reported that adipogenesisis a complex process which is tightly regulated by sequentialactivations of various transcriptional factors. Preadipocyte culturesystems are well established for studying cellular and molecularmechanisms of adipocyte differentiation. Adipokines, such as C/EBP α andPPARγ, are some of the most important genes during adipogenesis and havea direct impact on the development of fat cells.

Conclusion

The present findings show that the inventive composition can scavengefree radicals, inhibit the enzymes responsible for lipid metabolism, andinhibit adipAPPogenic differentiation of 3T3L-1 cells. These findingsdemonstrate that the composition can modulate molecular events in 3T3-L1 preadipocytes and thus provide a treatment for obesity.

1. A method of promoting a healthy body weight in a subject, comprising administering to the subject a composition comprising an effective amount of green tea flower extract.
 2. The method of claim 1, wherein said extract has an IC50 value of about 120 μg/mL for pancreatic lipase in vitro.
 3. The method of claim 1, wherein said extract has an IC50 value of about 199 μg/mL for alpha amylase in vitro.
 4. The method of claim 1, wherein said extract has an IC50 value of about 43 μg/mL for lipid peroxidation in vitro.
 5. The method of claim 1, wherein said extract has an IC50 of about 41 μg/mL as determined by DPPH radical scavenging assay.
 6. The method of claim 1, wherein administering said composition reduces body weight in said subject.
 7. The method of claim 1, wherein administering said composition prevents or inhibits weight gain in said subject.
 8. The method of claim 1, wherein administering said composition maintains said subject's body weight.
 9. The method of claim 1, wherein administering said composition reduces body fat in said subject.
 10. The method of claim 1, wherein administering said composition prevents or inhibits body fat gain in said subject.
 11. The method of claim 1, wherein said subject is overweight.
 12. The method of claim 1, wherein said subject is obese.
 13. The method of claim 1, wherein said extract is administered at a dose of about 50 mg per kilogram of bodyweight, about 100 mg per kilogram of bodyweight, about 150 mg per kilogram of bodyweight, or combinations thereof.
 14. (canceled)
 15. The method of claim 1, wherein said composition is administered daily, weekly, monthly, or combinations thereof.
 16. The method of claim 1, wherein said composition is administered in a form selected from the group consisting of a powder, liquid, pill, tablet, pellet, granule, capsule, soluble film, solution, spray, syrup, linctus, lozenge, pastille, chewing gum, chew, paste, vapor, suspension, emulsion, ointment, cream, lotion, foam, liniment, gel, drop, topical patch, buccal patch, bead, gummy, gel, sol, injection, and combinations thereof.
 17. The method of claim 1, wherein said composition further comprises at least one of a vitamin, mineral, amino acid, protein, carbohydrate, lipid, fatty acid, excipient, pharmaceutical carrier, bulking agent, binding agent, caffeine, flavoring, sweetener, and preservative.
 18. The method of claim 1, wherein said composition is selected from the group consisting of a food, snack, nutritional supplement, dietary supplement, food supplement, beverage, and combination thereof.
 19. The method of claim 1, wherein said composition is administered systemically.
 20. The method of claim 1, wherein said composition is administered by a route selected from the group consisting of orally, buccally, sub-lingually, and combinations thereof.
 21. (canceled)
 22. The method of claim 1, wherein said subject is a human. 